Wyss, Marianne; Gradauskaite, Vaiva; Ebert, Nadine; Thiel, Volker; Zurbriggen, Andreas; Plattet, Philippe (2022). Efficient Recovery of Attenuated Canine Distemper Virus from cDNA. Virus research, 316, p. 198796. Elsevier 10.1016/j.virusres.2022.198796
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1-s2.0-S016817022200123X-main.pdf - Published Version Available under License Creative Commons: Attribution (CC-BY). Download (2MB) | Preview |
To provide insights into the biology of the attenuated canine distemper virus (CDV) Onderstepoort (OP) strain (large plaque forming variant), design next-generation multivalent vaccines, or further investigate its promising potential as an oncolytic vector, we employed contemporary modifications to establish an efficient OP-CDV-based reverse genetics platform. Successful viral rescue was obtained however only upon recovery of a completely conserved charged residue (V13E) residing at the N-terminal region of the large protein (L). Although L-V13 and L-V13E did not display drastic differences in cellular localization and physical interaction with P, efficient polymerase complex (P+L) activity was recorded only with L-V13E. Interestingly, grafting mNeonGreen to the viral N protein via a P2A ribosomal skipping sequence (OPneon) and its derivative V-protein-knockout variant (OPneon-Vko) exhibited delayed replication kinetics in cultured cells. Collectively, we established an efficient OP-CDV-based reverse genetics system that enables the design of various strategies potentially contributing to veterinary medicine and research.
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